Abstract
Hematopoietic stem cell transplantation, which requires accurate enumeration of stem cells, is routinely used in clinical settings. Flow cytometry provides a qualitative and quantitative assessment of CD34⁺ cells. Precision, linearity, and stability of the novel BD™ Stem Cell Enumeration (SCE) Kit were evaluated on two flow cytometry platforms using a modified ISHAGE gating strategy and including a viability dye for data acquisition and analysis. Precision and linearity were evaluated on BD FACSCanto™ II and BD FACSCalibur™ systems. Stability was evaluated on the BD FACSCanto II system. Precision was tested using both high and low controls. Linearity was evaluated using dilutions from CD34⁺ cell pools, while stability was evaluated using fresh leukapheresis specimens. Both systems showed precision with limited variability in absolute counts and percentages of viable CD34⁺ cells. The linearity range of viable CD34⁺ cells in both systems was established at 0-1000 cells/μL, showing a linear relationship (R² = 0.99). Stability of CD34⁺ cells in mobilized leukapheresis samples was confirmed up to 24 h after collection and up to 60 min after the end of stain/lyse procedures. The BD SCE Kit on both flow cytometry systems shows consistent and reproducible results.
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